摘要
为探究绿原酸(CGA)对脂多糖(LPS)致急性肝损伤(ALI)大鼠铁死亡的影响及其对核因子E2相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)信号通路的调控作用,本试验选取32只雄性SD大鼠随机均分为CON组、CON+CGA组、LPS组和LPS+CGA组。CON组未作处理,LPS组腹腔注射10 mg/(kg·bw)LPS,CON+CGA组和LPS+CGA组均腹腔注射20 mg/(kg·bw)CGA,1 h后LPS+CGA组腹腔注射10 mg/(kg·bw)LPS。注射LPS 4 h后处死所有大鼠,采集血液和肝脏。测定血清谷草转氨酶(AST)和谷丙转氨酶(ALT)水平,苏木精-伊红(H.E.)染色观察肝脏病理学变化,测定肝脏氧化应激指标和亚铁离子(Fe^(2+))含量,普鲁士蓝染色观察铁沉积情况,实时荧光定量PCR(qPCR)检测铁死亡相关基因表达,蛋白免疫印迹法(Western blot)检测Nrf2和铁死亡相关蛋白表达,免疫组织化学法检测Nrf2蛋白表达和核转位情况。结果显示,CGA可极显著改善肝脏功能(P<0.01),减轻肝脏病理学损伤,抑制肝脏棕黄色铁颗粒沉积,极显著降低肝脏氧化应激损伤和Fe^(2+)含量(P<0.01),下调转铁蛋白受体1(TFR1)和二价金属离子转运蛋白1(DMT1)基因相对表达量(P<0.01),上调GPX4基因相对表达量和Nrf2、溶质载体家族7成员11(SLC7A11)、GPX4蛋白表达量(P<0.01)。结果表明,CGA通过激活Nrf2/GPX4信号通路,抑制肝脏脂质过氧化和铁死亡,改善肝脏功能,从而缓解LPS诱导的大鼠ALI。本试验为兽医临床防治脓毒症致ALI提供了潜在治疗靶点和新思路。
This study aimed to explore the effect of chlorogenic acid(CGA)on ferroptosis in rats with acute liver injury(ALI)induced by lipopolysaccharide(LPS),and its regulatory role on the nuclear factor erythroid 2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)signaling pathway.Thirty-two male SD rats were randomly divided into four groups:control(CON),CON+CGA,LPS,and LPS+CGA.The CON group received no treatment;the LPS group received an intraperitoneal injection of LPS[10 mg/(kg·bw)];the CON+CGA and LPS+CGA groups were pretreated with CGA[20 mg/(kg·bw)]via intraperitoneal injection;one hour later,the LPS+CGA group received LPS[10 mg/(kg·bw)]injection.All rats were euthanized 4 hours after LPS administration,and blood and liver samples were collected.Serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were measured to assess liver function.Liver pathological changes were observed via hematoxylin-eosin(H.E.)staining.Oxidative stress indicators and ferrous iron(Fe^(2+))content were determined.Prussian blue staining was used to assess hepatic iron deposition.Real-time fluorescent quantitative PCR(qPCR)was conducted to analyze expression of ferroptosisrelated genes,and Western blot was used to evaluate the expression of Nrf2 and ferroptosis-related proteins.Immunohistochemistry was employed to detect Nrf2 expression and nuclear translocation.The results showed that CGA significantly improved liver function(P<0.01),alleviated pathological liver damage,inhibited hepatic iron deposition,and reduced oxidative stress and Fe^(2+)levels in the liver(P<0.01).Furthermore,CGA downregulated the mRNA levels of transferrin receptor 1(TFR1)and divalent metal ion transporter 1(DMT1)(P<0.01),while upregulating the mRNA expression of GPX4 and protein levels of Nrf2,solute carrier family 7 member 11(SLC7A11),and GPX4(P<0.01).These findings indicate that CGA exerts protective effects against LPS-induced ALI in rats by activating the Nrf2/GPX4 signaling pathway,thereby suppressing lipid peroxidation and ferroptosis in the liver.This study provides potential therapeutic targets and novel strategies for veterinary clinical management of sepsis-induced acute liver injury.
作者
黄静
王铭铭
刘敬轩
淳海清
刘恩喜
唐琦超
HUANG Jing;WANG Mingming;LIU Jingxuan;CHUN Haiqing;LIU Enxi;TANG Qichao(College of Animal Science and Technology,Jilin Agricultural Science and Technology University,Jilin 132101,China)
出处
《中国兽医杂志》
北大核心
2025年第8期37-45,共9页
Chinese Journal of Veterinary Medicine
基金
吉林农业科技学院国家级大学生科技创新创业训练计划项目(GJ202211439006)
深圳市瑞鹏公益基金会与新瑞鹏宠物医疗集团有限公司共同资助高校青年教师科研基金项目(RPJJ2021027)。
